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The need for finding fungicides against  Fusarium  is a key step in the chemical plant protection and using appropriate chemical agents. Existing, conventional methods of evaluation of  Fusarium  isolates resistance to fungicides are costly, time-consuming and potentially environmentally harmful due to usage of high amounts of potentially toxic chemicals. Therefore, the development of fast, accurate and effective detection methods for  Fusarium  resistance to fungicides is urgently required. MT2 microplates (Biolog TM ) method is traditionally used for bacteria identification and the evaluation of their ability to utilize different carbon substrates. However, to the best of our knowledge, there is no reports concerning the use of this technical tool to determine fungicides resistance of the  Fusarium  isolates. For this reason, the objectives of this study are to develop a fast method for  Fusarium  resistance to fungicides detection and to validate the effectiveness approach between both traditional hole-plate and MT2 microplates assays. In presented study MT2 microplate-based assay was evaluated for potential use as an alternative resistance detection method. This was carried out using three commercially available fungicides, containing following active substances: triazoles (tebuconazole), benzimidazoles (carbendazim) and strobilurins (azoxystrobin), in six concentrations (0, 0.0005, 0.005, 0.05, 0.1, 0.2%), for nine selected  Fusarium  isolates. In this study, the particular concentrations of each fungicides was loaded into MT2 microplate wells. The wells were inoculated with the  Fusarium  mycelium suspended in PM4-IF inoculating fluid. Before inoculation the suspension was standardized for each isolates into 75% of transmittance. Traditional hole-plate method was used as a control assay. The fungicides concentrations in control method were the following: 0, 0.0005, 0.005, 0.05, 0.5, 1, 2, 5, 10, 25, and 50%. Strong relationships between MT2 microplate and traditional hole-plate methods were observed regarding to the detection of  Fusarium  resistance to various fungicides and their concentrations. The tebuconazole was most potent, providing increased efficiency in the growth inhibition of all tested isolates. Almost all among tested isolates were resistant to azoxystrobin-based fungicide. Overall, the MT2 microplates method was effective and timesaving, alternative method for determining  Fusarium  resistance/sensitivity to fungicides, compering to traditional hole-plate approach.

Fast and Accurate Microplate Method (Biolog MT2) for Detection of Fusarium Fungicides Resistance/Sensitivity